Independent, Controllable Stretch-Perfusion Bioreactor Chambers to Functionalize Cell-Seeded Decellularized Tendons

Giuseppe Talò,Matteo Moretti,Sergio Lorenzi,Daniele D’Arrigo,Arianna B Lovati

doi:10.1007/s10439-019-02257-6

Abstract

Tissue-engineered decellularized matrices can progress clinical replacement of full-thickness ruptures or tendon defects. This study develops and validates a custom-made automated bioreactor, called oscillating stretch-perfusion bioreactor (OSPB), consisting of multiple, independent culture chambers able to combine a bidirectional perfusion with a programmable, uniaxial strain to functionalize cell-seeded decellularized tendons. Decellularized tendon matrices were seeded on their surfaces and within the tendon fibers with mesenchymal stem cells. Then, they were subjected to a bidirectional perfusion and programmed stretching cycles of 15–30–60 min on–off two times per day for 7 days of culture. In vitro analyses showed viable cells, homogenously distributed on the surface of the constructs. More importantly, cell-seeded decellularized tendon grafts undergoing cyclic load in our bioreactor had a superior production and organization of newly formed collagen matrix compared to static cultured constructs. The coherency and local alignment of the new collagen deposition within the inner injected channels quantitatively supported histological findings. The designed OSPB could be considered a unique, cost-effective system able to involve multiple independently controlled chambers in terms of biological and mechanical protocols. This system allows parallel processing of several customized tendon constructs to be used as grafts to enhance the surgical repair of large tendon defects.

Highlights

Tendon injuries and full-thickness ruptures represent common occurrences in musculoskeletal disorders, affecting patients’ quality of life and treatment costs
The device was developed following main requirements: (i) independent chambers that fit on an oscillating platform exploiting perfusion and medium oxygenation working principles; (ii) sterilizable, biocompatible and chemically inert chamber materials; (iii) easy removable culture chamber from the bioreactor to be handled under laminar flow hoods; (iv) chambers provide a tissue holder consisting of a clamping grip to safely block the tendon without damaging the tissue; (v) the holder accommodates samples with clinically relevant dimensions; (vi) each chamber provides an axial stretching to the constructs with a preload and physiological-like stress/strain that could be programmed, real-time controlled and monitored during culture through a feedback loop control; (vii) the entire bioreactor system must be accommodated into standard cell culture incubators
The purpose of this study was the development of a bioreactor to dynamically culture biological constructs for the replacement of tendon defects

Summary

Introduction

Tendon injuries and full-thickness ruptures represent common occurrences in musculoskeletal disorders, affecting patients’ quality of life and treatment costs. With a low metabolic activity, tendons have a sub-optimal capability to heal with enhanced risks of re-injuries. In these cases, graft replacement is the gold standard approach. Among several types of grafts, research is focused on decellularized matrices derived from humans or animals to better resemble complex biochemical, ultrastructural and mechanical properties of native tendons.. A good removal of cell-related immunogenicity of these grafts can be obtained through decellularization protocols.. A good removal of cell-related immunogenicity of these grafts can be obtained through decellularization protocols.2,8,29 These biological, non-vital implants are prone to degeneration and limited duration after Among several types of grafts, research is focused on decellularized matrices derived from humans or animals to better resemble complex biochemical, ultrastructural and mechanical properties of native tendons. A good removal of cell-related immunogenicity of these grafts can be obtained through decellularization protocols. these biological, non-vital implants are prone to degeneration and limited duration after

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